I am using Invitrogen's Fluo4 assay to measure intracellular calcium levels. I load my cells with the dye for an hour, then measure the fluorescence with a flow cytometer for a minute before I add thapsigargin to release ER calcium stores and then measure fluorescence for another 4 minutes.
My fluroescence vs. time plot has a peak after I add the thapsigargin, which I would expect because calcium is released and binds to the indicator, but then the fluroescence decreases after that (after about a minute after thapsigargin was added). I was just wondering why the fluroescence would taper off...???....I think this is also seen in other dyes.
Thanks for any insights in advance!
calcium flux measurement
No replies to this topic